HPV16 L2肽段疫苗的构建及免疫学功能研究

Construction and immunizing potency of HPV16 L2 peptiede vaccine

目的探讨基于人乳头瘤病毒16次要衣壳蛋白L2抗原优势表位多倍复制重组多肽疫苗的构建与诱导免疫应答之间的关系。方法采用芴甲氧羰基（Fmoc）固相合成法合成多肽，用高效液相色谱分析多肽的纯度，对所合成的多肽采用质谱进行定性鉴定。通过淋巴细胞增殖试验、细胞毒性杀伤试验及酶联免疫吸附试验方法研究该疫苗激发的细胞免疫及反应强度。结果合成的多肽经纯化后纯度均达到95％以上，经质谱分析，各肽的分子量测定值与理论值相符。所设计的多肽疫苗免疫小鼠后，小鼠淋巴细胞体外培养增殖明显，并可特异性地在体外杀伤小鼠肺上皮细胞，小鼠血清体外可诱导较高的干扰素-γ。结论在多肽疫苗的设计中，多倍复制重组人乳头瘤病毒次要衣壳蛋白L2肽段优势表位可增强疫苗的免疫原性，为进一步研制开发宫颈癌疫苗打下了基础。

Objective To explore relationship between construction of dominant epitope human papillomavirus 16 （HPV16） minor eapsid protein L2 based polyduplicated recombinant polypeptide vaccine and induced immunologic response. Methods Fmoc solid-phase synthesis was used to synthesize polypeptide, purity of the polypeptide was analyzed by using high performance liquid chromatography and the synthesized polypeptides were identified qualitatively by using spectrometry. The vaccine-induced cellular immunity and response intensity were examined by using lymphocyte proliferation test, cell eytotoxieity test and ELISA method. Results After purification, the purity of synthesized polypeptides all reached over 95%. The mass spectrometry analysis showed the measured values of each peptide molecular weight were in accordance with the theoretical value . After being immunized by designed polypeptide vaccine, proliferation of murine lymphocytes in culture in vitro was evident and TC-1 cells （ lung epithelial cells of mice） could be killed specifically by the murine lymphocytes in vitro. Mice serum could induce high concentration of IFN-γ in vitro. Conclusion In design of polypeptide vaccine, polyduplication and rearrangement of human papillomavirus 16 minor eapsid protein L2 peptide dominant epitopes can enhance the vaccine＇ s immunogenicity, which lays a solid foundation for future research and development of vaccine for cervical carcinoma.